The aim of the present study was to determine whether clinical or laboratory factors can influence the development of single pronucleated zygotes (1PN) and two polar bodies (PB) after ICSI. In total, 341 ICSI cycles performed at FertiClinic-Villa Margherita from January 2012 to December 2014 were enrolled in the study. Group A included 240 cycles with no 1PN−2PB while group B included 101 cycles with one or more 1PN−2PB. Age, stimulation protocol, infertility factor, amount of gonadotropin administered, duration of therapy, peak estradiol levels, number of follicles at maturation triggering, oocytes retrieved and mature oocytes, time between retrieval and injection and sperm characteristics were compared between groups. In opposition to previous results showing no relationship between 1PN occurrence and clinical or laboratory variables, we observed that 1PN−2PB zygote formation seems to be associated with a lower female age, higher level of E2 and higher number of follicles on day of oocyte maturation triggering, higher number of astenozoospermic male patients, more oocytes retrieved at pick-up, more mature oocytes (MII) and longer time to injection.

The present study aimed to determine the relationship among changes in the number of preantral follicles and concentrations of follicle-stimulating hormone (FSH), luteinizing hormone (LH), progesterone (P4), androstenedione (A) and estradiol-17β (E2) in the fetal heart, umbilical cord and maternal blood. Primordial follicles had already appeared in a 20 cm fetus and primary follicles were observed in a 50 cm fetus. In a 70 cm fetus, the number of primordial and primary follicles increased rapidly and secondary follicles were present. The concentrations of LH and FSH did not change between 20 cm and 160 cm in fetal length. When the fetal length became >70 cm, serum levels in the fetus, umbilical cord and mothers, and E2 levels in umbilical cord increased synchronously (p<0.05). These results showed increases in the number of preantral follicles in the Antarctic minke whale fetal ovary along with fetal growth during the early gestation period. These findings suggest that the change in preantral follicles was associated with changes in the concentration of steroids in early gestation periods. The changes in steroid concentrations in the fetal and umbilical cord blood and the increased number of preantral follicles were coincident at around 70 cm in fetal length, whereas the growth and differentiation of primordial and primary follicles appeared to be independent of FSH and LH.

Calcium signalling is involved in important events in oocytes, such as meiotic competence acquisition. We have previously demonstrated the positive influence of animal age and gonadotropin stimulation in vivo regarding the ability of oocytes recovered from preantral follicles to exhibit calcium spikes. In the present work we determined whether preantral follicle development in vitro also allows oocytes to acquire calcium signalling activity. We also aimed to verify the influence of animal age, FSH + LH and/or insulin on oocyte calcium spike acquisition during preantral follicle culture. Early preantral follicles were isolated from 12-day-old and 1- to 3-month-old F1 hybrid mice and cultured individually for either 2 or 6 days. At the end of the culture period the oocytes were processed for calcium imaging by confocal microscopy. We show that oocytes recovered from cultured preantral follicles exhibit variable calcium spike activity rates, depending on animal age, culture duration and hormonal supplementation. Oocytes recovered from adult animals continue to exhibit calcium spikes, and those recovered from juveniles acquire that activity after culture. Insulin and gonadotropins in combination account for an early and maintained inhibitory effect on calcium signalling acquisition by oocytes. Insulin alone also leads to an early inhibitory effect, which, however, disappears with longer culture periods. Contrary to the complex in vivo situation, the acquisition of calcium signalling by oocytes in a controlled in vitro environment does not seem to be dependent on gonadotropins alone.

The main objective of the experiment was to investigate the mechanism by which progesterone priming eliminates defective luteal function in anoestrous ewes induced to ovulate with GnRH. Animals in group 1 (no. = 10) were primed with a single i.m. injection of progesterone in corn oil 3 days before the start of GnRH treatment, while ewes in group 2 (no. = 10) received corn oil alone and served as untreated controls. Ewes in group 3 (no. = 10), which served as positive controls, were treated with an intra-vaginal progestagen sponge for 7 days, and this was removed just before the start of GnRH treatment. Ewes in all three groups were induced to ovulate by administration of 2-h injections of GnRH (250 ng per injection) for 54 h. Frequent blood samples for LH, FSH and progesterone analysis were taken around the time of both progesterone injection and GnRH treatment, as well as daily thereafter to monitor luteal function, and laparoscopy was performed 3 and 7 days after GnRH treatment.

The incidence of ovulation was similar for all the three groups (8/10, 7/10 and 9/10 for groups 1, 2 and 3 respectively). Hoioever, both laparoscopic examination and plasma progesterone concentrations revealed that the incidence of normal luteal function was significantly higher in progesterone-primed animals group 1:7/8; group 3:9/9) compared ivith controls (group 2:0/7), P < 0-05) with no difference between groups 1 and 3. Injection of progesterone on day −3 significantly suppressed mean LH concentrations (P < 0·05), but mean FSH concentrations were not altered. However, there were no significant differences between groups in LH and FSH concentrations over the period of GnRH treatment, nor in the timing, duration and height of pre-ovulatory LH and FSH surges. These results suggest that progesterone priming may eliminate defective luteal function either by changing LH concentrations at the time of progesterone administration or through mechanisms not involving gonadotropin secretion.

One of the more promising techniques for the improvement of sow productivity has been the use of partial weaning or the separation of the sow and litter for periods of between 3 to 12 h/day towards the end of lactation. The purpose of the present study was to examine changes in steroid hormone concentrations, oestrus and fertility responses in lactating sows subjected to altered sucking, boar exposure and gonadotropins in mid lactation and at weaning. Thirty-two crossbred sows were allocated at random to one of four experimental treatments. Treatment 1 sows (no. = 8) were separated from their respective litters for 3 h/day from 12 days post partum through until weaning which was carried out at 21 days post partum. Treatment 1 sows were also given in-pen boar exposure for 1 h/day between day 12 post partum and weaning. Treatment 2 sows (no. = 8) were given the same protocol of litter separation (LS) and boar exposure (BE) as in treatment 1 and they were also given a combination of gonadotropins (Gn) on day 17 of lactation. Treatment 3 sows (no. = 8) were given LS + BE as above and the sows were injected with Gn on the day of weaning. Treatment 4 sows were controls given no LS, BE or Gn (no. = 8). During lactation, treatment 1 sows exhibited significantly (P < 0·05) reduced progesterone concentrations compared with treatment 2 sows. There were no other significant effects of treatment for any of the periods considered. The separation of sows from their piglets with or without gonadotropic treatment did not result in lactational oestrus in any treated sows. No significant treatment differences were found in the subsequent litter size (piglets born alive or total number of piglets). In conclusion, the present results have demonstrated no significant effects of LS, BE and Gn treatment during lactation on sow and litter performance. This was associated with low levels of peripheral plasma progesterone in all lactating sows.

The effect of body condition on hypothalamic and pituitary function in post-partum suckled beef cows was studied. Thirty-eight Blue-Grey cows were calved at a body condition score of either 2·35 (L) or 2·80 (H), fed to maintain live weight and body condition thereafter and ovariectomized at either 5 or 9 weeks post partum. The H cows had higher LH pulse frequencies before ovariectomy (0·36 v. 0·28 pulses per h; P < 0·05) and 15 days after ovariectomy (0·86 v. 0·72 pulses per h; P < 0·01), indicating a direct effect of body condition on GnRH release from the hypothalamus. Body condition had no effect on the ability of the pituitary to release LH in response to a physiological dose (2·5 u.g) of GnRH in entire or ovariectomized cows at 5 or 9 weeks post partum. The magnitude of the increase in circulating levels of LH following GnRH injection was greater at 9 weeks compared with 5 weeks post partum (2·23 v. 1·73 u.g/1; P < 0·001) in entire cows, but there was no difference between 5 and 9 weeks in ovariectomized cows. Following ovariectomy there was a 2·5-fold increase in LH pulse frequency within 15 days and LH pulse frequency approached 1 pulse per h in all treatments groups. It is concluded that both the hypothalamus and the pituitary are capable of functioning normally by 5 weeks post partum in suckled beef cows, that the pulsatile release of GnRH from the hypothalamus is inhibited, possibly due to a very high sensitivity to oestradiol, and that the sensitivity of the pituitary to the negative feedback effect of oestradiol may decrease with time post partum.

A total of 178 placentas of twins born in Tokyo in 1961-1982 were examined. Only just over one third were dichorionic, 110 being monochorionic and DZ twins were estimated to account for 24.7% of the pairs. The lower frequency of DZ twinning in Japan is attributed to lower gonadotropin values in Japanese women as compared to European, American, or African women. High fetal morbidity was found in monochorionic twins and this is attributed to a frequent occurrence of vascular anastomoses.

Serum gonadotropin, estradiol, prolactin and alpha-1-antitrypsin levels were measured in the first four days of the menstrual cycle in 14 women who were cycling normally. FSH, and to a lesser extent LH levels, were significantly higher in a group of 8 women who had at least one set of DZ twins (6 of whom had 2 sets) than in a control group of 6 women with no DZ twins. Estradiol levels also tended to be higher in mothers of twins but there were no significant differences in prolactin or alpha-1-antitrypsin concentrations. There were no differences between the means of the two groups in age, height, weight or number of pregnancies.

A follow-up of 78 twin pregnancies diagnosed in the first trimester indicated that early ovular resorption occurred in 63% of spontaneous and 64% of Clomiphene-induced gestations, so that singletons were born. No failure was observed in the 12 twin conceptions following administration of gonadotropins or Cyclofenil.